We found a match
Your institution may have rights to this item. Sign in to continue.
- Title
Purification of a Low-Molecular-Weight Phospholipase A2 Associated with Soluble High-Molecular-Weight Acidic Proteins from Rabbit Nucleus Pulposus and Its Comparison with a Rabbit Splenic Group IIa Phospholipase A21.
- Authors
Tanaka, Naruhiro; Ishida, Tetsuo; Hukuda, Sinsuke; Horiike, Kihachiro
- Abstract
An intervertebral disc is a large peice of avascular cartilage rich in proteoglycans and water consisting of gelatinous nucleus pulposus and fibrous annulus fibrosus. The soluble fraction of rabbit nucleus pulposus exhibited unusually high Ca2+-dependent phospholipase A2 (PLA2) activity (about 70% of the total PLA2 activity). The soluble PLA2 activity was 6-7-fold higher than those of rabbit annulus fibrosus and spleen. The PLA2 was bound to an anion-exchange column at pH 7.4, and eluted near the void volume as a broad peak on gel-filtration on a TSKgel SuperSW3000 column developed with a buffer containing 0.1–0.2 M salt When the gel-filtration column was developed in the presence of 1 M salt, almost all the PLA2 activity was eluted near the total available volume. The soluble PLA2 was purified to near homogeneity. A Ca2+-dependent PLA2 was also purified from the fractions extracted with 1 M KBr from nucleus pulposus. For comparison, we purified a Ca2+-dependent PLA2 from the KBr fraction of spleen. The splenic PLA2 was identical to a group IIa PLA2, as judged from its N-terminal amino acid sequences and mass spectra. On SDS—polyacrylamide gel electrophoresis the enzymes purified from the soluble and KBr fractions of nucleus pulposus both gave a major 15.7-kDa band at the same position as splenic group IIa PLA2. These results suggest that group IIa PLA2 is associated with soluble high-molecular-weight proteins, most likely proteoglycans, in the extracellular matrix of rabbit nucleus pulposus.
- Subjects
PHOSPHOLIPASES; PROTEINS; AMINO acids; NUCLEUS pulposus; SPLENIC rupture
- Publication
Journal of Biochemistry, 2000, Vol 127, Issue 6, p985
- ISSN
0021-924X
- Publication type
Article
- DOI
10.1093/oxfordjournals.jbchem.a022715